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Study on Enhancement Principle and Stabilization for the Luminol-H2O2-HRP Chemiluminescence System.

Yang L, Jin M, Du P, Chen G, Zhang C, Wang J, Jin F, Shao H, She Y, Wang S, Zheng L, Wang J - PLoS ONE (2015)

Bottom Line: First, the comparative study on the enhancement effect of ten compounds as enhancers to the luminol-H2O2-HRP chemiluminescence system was carried out, and the results showed that 4-(imidazol-1-yl)phenol (4-IMP), 4-iodophenol (4-IOP), 4-bromophenol (4-BOP) and 4-hydroxy-4'-iodobiphenyl (HIOP) had the best performance.When the concentration of HRP ranges from 6 to 25 ng/mL, 4-IOP was the best choice.Finally, the three well-performing chemiluminescent enhanced solutions (CESs) have been further optimized according to the three enhancers (4-IMP, 4-IOP and HIOP) in their utilized HRP concentration ranges.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.

ABSTRACT
A luminol-H2O2-HRP chemiluminescence system with high relative luminescent intensity (RLU) and long stabilization time was investigated. First, the comparative study on the enhancement effect of ten compounds as enhancers to the luminol-H2O2-HRP chemiluminescence system was carried out, and the results showed that 4-(imidazol-1-yl)phenol (4-IMP), 4-iodophenol (4-IOP), 4-bromophenol (4-BOP) and 4-hydroxy-4'-iodobiphenyl (HIOP) had the best performance. Based on the experiment, the four enhancers were dissolved in acetone, acetonitrile, methanol, and dimethylformamide (DMF) with various concentrations, the results indicated that 4-IMP, 4-IOP, 4-BOP and HIOP dissolved in DMF with the concentrations of 0.2%, 3.2%, 1.6% and 3.2% could get the highest RLU values. Subsequently, the influences of pH, ionic strength, HRP, 4-IMP, 4-IOP, 4-BOP, HIOP, H2O2 and luminol on the stabilization of the luminol-H2O2-HRP chemiluminescence system were studied, and we found that pH value, ionic strength, 4-IMP, 4-IOP, 4-BOP, HIOP, H2O2 and luminol have little influence on luminescent stabilization, while HRP has a great influence. In different ranges of HRP concentration, different enhancers should be selected. When the concentration is within the range of 0~6 ng/mL, 4-IMP should be selected. When the concentration of HRP ranges from 6 to 25 ng/mL, 4-IOP was the best choice. And when the concentration is within the range of 25~80 ng/mL, HIOP should be selected as the enhancer. Finally, the three well-performing chemiluminescent enhanced solutions (CESs) have been further optimized according to the three enhancers (4-IMP, 4-IOP and HIOP) in their utilized HRP concentration ranges.

No MeSH data available.


Related in: MedlinePlus

The effect of organic solvent concentration on the CL intensity.a) Luminol-H2O2-HRP-HIOP, b) Luminol-H2O2-HRP-4-IOP, c) Luminol-H2O2-HRP-4-BOP, d) Luminol-H2O2-HRP-4-IMP.
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pone.0131193.g002: The effect of organic solvent concentration on the CL intensity.a) Luminol-H2O2-HRP-HIOP, b) Luminol-H2O2-HRP-4-IOP, c) Luminol-H2O2-HRP-4-BOP, d) Luminol-H2O2-HRP-4-IMP.

Mentions: HIOP dissolved by DMF has better luminescence-enhancing effects compared with HIOP dissolved by acetone, with an enhanced luminescence value reaching 109. The luminescence value will increase along with the increase of DMF content (see Fig 2a and S2 Text), which may be because HIOP is difficult to dissolve in water, and will separate out a white precipitate in a hydrofacies luminescence system. As Fig 2b and S3 Text demonstrate, the order of magnitude of 4-IOP enhanced luminescence reaches 109; the enhancement effects of DMF are relatively good, while the enhancement effects of acetonitrile are the least. The influence of organic solvent in terms of luminescence-enhancing effects is very small. As Fig 2c and S4 Text show, when the content of DMF in the luminescence system reaches 1.6%, the dissolved 4-BOP enhancement effect is best and luminescence is at the maximum value. Along with the increase of acetone content, the enhanced luminescence of 4-BOP is increased too, with its luminescence value reaching 109. When using DMF to dissolve 4-IMP, the enhanced luminescence will reach 109; when the content of DMF is greater than 10%, the effect decreases (see Fig 2d and S5 Text). As in summary, if the concentration of organic solvents in the buffer was too low, the enhancer may be precipitated in the buffer system and the RLU will at the low level. And if the concentration of organic solvents in the buffer was too high, it could affect the activity of the HRP and decrease the RLU. So we thought the combination of two factors mentioned above was the key factor how the concentrations of organic solvents affect the RLU of the chemiluminscent system.


Study on Enhancement Principle and Stabilization for the Luminol-H2O2-HRP Chemiluminescence System.

Yang L, Jin M, Du P, Chen G, Zhang C, Wang J, Jin F, Shao H, She Y, Wang S, Zheng L, Wang J - PLoS ONE (2015)

The effect of organic solvent concentration on the CL intensity.a) Luminol-H2O2-HRP-HIOP, b) Luminol-H2O2-HRP-4-IOP, c) Luminol-H2O2-HRP-4-BOP, d) Luminol-H2O2-HRP-4-IMP.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495922&req=5

pone.0131193.g002: The effect of organic solvent concentration on the CL intensity.a) Luminol-H2O2-HRP-HIOP, b) Luminol-H2O2-HRP-4-IOP, c) Luminol-H2O2-HRP-4-BOP, d) Luminol-H2O2-HRP-4-IMP.
Mentions: HIOP dissolved by DMF has better luminescence-enhancing effects compared with HIOP dissolved by acetone, with an enhanced luminescence value reaching 109. The luminescence value will increase along with the increase of DMF content (see Fig 2a and S2 Text), which may be because HIOP is difficult to dissolve in water, and will separate out a white precipitate in a hydrofacies luminescence system. As Fig 2b and S3 Text demonstrate, the order of magnitude of 4-IOP enhanced luminescence reaches 109; the enhancement effects of DMF are relatively good, while the enhancement effects of acetonitrile are the least. The influence of organic solvent in terms of luminescence-enhancing effects is very small. As Fig 2c and S4 Text show, when the content of DMF in the luminescence system reaches 1.6%, the dissolved 4-BOP enhancement effect is best and luminescence is at the maximum value. Along with the increase of acetone content, the enhanced luminescence of 4-BOP is increased too, with its luminescence value reaching 109. When using DMF to dissolve 4-IMP, the enhanced luminescence will reach 109; when the content of DMF is greater than 10%, the effect decreases (see Fig 2d and S5 Text). As in summary, if the concentration of organic solvents in the buffer was too low, the enhancer may be precipitated in the buffer system and the RLU will at the low level. And if the concentration of organic solvents in the buffer was too high, it could affect the activity of the HRP and decrease the RLU. So we thought the combination of two factors mentioned above was the key factor how the concentrations of organic solvents affect the RLU of the chemiluminscent system.

Bottom Line: First, the comparative study on the enhancement effect of ten compounds as enhancers to the luminol-H2O2-HRP chemiluminescence system was carried out, and the results showed that 4-(imidazol-1-yl)phenol (4-IMP), 4-iodophenol (4-IOP), 4-bromophenol (4-BOP) and 4-hydroxy-4'-iodobiphenyl (HIOP) had the best performance.When the concentration of HRP ranges from 6 to 25 ng/mL, 4-IOP was the best choice.Finally, the three well-performing chemiluminescent enhanced solutions (CESs) have been further optimized according to the three enhancers (4-IMP, 4-IOP and HIOP) in their utilized HRP concentration ranges.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.

ABSTRACT
A luminol-H2O2-HRP chemiluminescence system with high relative luminescent intensity (RLU) and long stabilization time was investigated. First, the comparative study on the enhancement effect of ten compounds as enhancers to the luminol-H2O2-HRP chemiluminescence system was carried out, and the results showed that 4-(imidazol-1-yl)phenol (4-IMP), 4-iodophenol (4-IOP), 4-bromophenol (4-BOP) and 4-hydroxy-4'-iodobiphenyl (HIOP) had the best performance. Based on the experiment, the four enhancers were dissolved in acetone, acetonitrile, methanol, and dimethylformamide (DMF) with various concentrations, the results indicated that 4-IMP, 4-IOP, 4-BOP and HIOP dissolved in DMF with the concentrations of 0.2%, 3.2%, 1.6% and 3.2% could get the highest RLU values. Subsequently, the influences of pH, ionic strength, HRP, 4-IMP, 4-IOP, 4-BOP, HIOP, H2O2 and luminol on the stabilization of the luminol-H2O2-HRP chemiluminescence system were studied, and we found that pH value, ionic strength, 4-IMP, 4-IOP, 4-BOP, HIOP, H2O2 and luminol have little influence on luminescent stabilization, while HRP has a great influence. In different ranges of HRP concentration, different enhancers should be selected. When the concentration is within the range of 0~6 ng/mL, 4-IMP should be selected. When the concentration of HRP ranges from 6 to 25 ng/mL, 4-IOP was the best choice. And when the concentration is within the range of 25~80 ng/mL, HIOP should be selected as the enhancer. Finally, the three well-performing chemiluminescent enhanced solutions (CESs) have been further optimized according to the three enhancers (4-IMP, 4-IOP and HIOP) in their utilized HRP concentration ranges.

No MeSH data available.


Related in: MedlinePlus